Abstract
Multidrug-resistant bacteria are a growing problem worldwide. One extensively studied resistance mechanism is biofilm colonization—microbial colonies formed by many Gram-positive and Gram-negative bacteria species. Cationic antimicrobial peptides (AMPs) are innate immune system molecules serving as a first line of defense in fighting invading pathogens. The AMPs’ underlying mechanism and biophysical properties required for anti-biofilm activity are not fully known. Here we present protocols for investigating AMPs’ biological activity against major stages of biofilm life cycle, namely, planktonic stage (MIC assay), initial adhesion to surfaces (bacterial attachment assay), and formation or degradation of sessile microcolonies (biofilm formation and degradation assays). Furthermore, we demonstrate experiments that allow determination and comparison between peptide biophysical properties (secondary structure, hydrophobicity, and oligomerization) and how they affect their mechanism (peptide-binding assays) of anti-biofilm activity.
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References
Parsek MR, Singh PK (2003) Bacterial biofilms: an emerging link to disease pathogenesis. Ann Rev Microbiol 57:677–701
de Kievit TR (2009) Quorum sensing in Pseudomonas aeruginosa biofilms. Environ Microbiol 11:279–288
Keller L, Surette MG (2006) Communication in bacteria: an ecological and evolutionary perspective. Nature Reviews Microbiol 4:249–258
Mah TF, O’Toole GA (2001) Mechanisms of biofilm resistance to antimicrobial agents. Trends in microbiol 9:34–39
Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284:1318–1322
Hall-Stoodley L, Costerton JW, Stoodley P (2004) Bacterial biofilms: from the natural environment to infectious diseases. Nature Reviews Microbiol 2:95–108
Jorge P, Lourenco A, Pereira MO (2012) New trends in peptide-based anti-biofilm strategies: a review of recent achievements and bioinformatic approaches. Biofouling 28:1033–1061
Rossi LM, Rangasamy P, Zhang J, Qiu XQ, Wu GY (2008) Research advances in the development of peptide antibiotics. J Pharmaceuti Sci 97:1060–1070
Lehrer RI, Ganz T (1999) Antimicrobial peptides in mammalian and insect host defence. Curr Opin Immunol 11:23–27
Shai Y (2002) Mode of action of membrane active antimicrobial peptides. Biopolymers 66:236–248
Hall-Stoodley L, Stoodley P (2005) Biofilm formation and dispersal and the transmission of human pathogens. Trends in microbiology 13:7–10
Overhage J, Campisano A, Bains M, Torfs EC, Rehm BH, Hancock RE (2008) Human host defense peptide LL-37 prevents bacterial biofilm formation. Infection Imm 76:4176–4182
Segev-Zarko L, Saar-Dover R, Brumfeld V, Mangoni ML, Shai Y (2015) Mechanisms of biofilm inhibition and degradation by antimicrobial peptides. Biochem J 468:259–270
Saar-Dover R, Bitler A, Nezer R, Shmuel-Galia L, Firon A, Shimoni E, Trieu-Cuot P, Shai Y (2012) D-alanylation of lipoteichoic acids confers resistance to cationic peptides in group B streptococcus by increasing the cell wall density. PLoS pathogens 8:e1002891
Acknowledgment
This study was supported by The Pasteur-Weizmann Foundation, Israel Science Foundation (ISF) and the German-Israel Foundation (GIF).
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Segev-Zarko, La., Shai, Y. (2017). Methods for Investigating Biofilm Inhibition and Degradation by Antimicrobial Peptides. In: Hansen, P. (eds) Antimicrobial Peptides. Methods in Molecular Biology, vol 1548. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6737-7_22
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DOI: https://doi.org/10.1007/978-1-4939-6737-7_22
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6735-3
Online ISBN: 978-1-4939-6737-7
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